Cell Preps

Preparations #

11/30/21: Chemically competent cells #

Inoculated 1L LB in a 4 L autoclaved flask with 5 ml DH10B cells from liquid culture grown at 37C and started at 7:30 am this morning. Inoculated 1 L LB at 5:35 pm and grow overnight at 19C second floor innova shaker.

12/1/21: Chemically competent cells continued #

Came into lab at 7:50 am to start recording OD of cells started yesterday.

OD readings #

OD reading are made against sample taken from LB before inoculating with live culture.

07:50:00 AM0.18
08:50:00 AM0.28
09:50:00 AM0.27
10:32:00 AM0.3
11:16:00 AM0.34
11:48:00 AM0.38
12:20:00 PM0.43

Placed cells on ice. Followed Al-basam provided lab protocol and dispensed cells into 0.5 ml tubes in 200 ul aliquots. Daisy and Megan dropped tubes into LN2 as they were filled. Stored samples in -80C freezer.

Competency test #

After freezing thawed 4 aliquots of cells to test efficiency using NEB protocol for transformations. Table below describes samples. Plated 1/10th total transformed volume.

SampleVolume plasmid (ul)DNA mass (ug)Plasmid

Incubated at 37C overnight.

PlateDNA mass (pg)Proportion platedNumber of ColoniesEfficiency